A Surveillance Study of Cooling Tower Water of Large Buildings in Seoul City for Legionella Species with Molecular Analysis

Kim, Kwon Beom; Kim, Woo Joo; Kim, Min Ja; Park, Seung Chul; You, Se Hwa; Shim, Hee Sun; Ham, Hee Jin; Park, Suk Gi

Abstract

BACKGROUND
To prevent and control legionellosis outbreaks, it is important to monitor cooling towers for Legionella and establish epidemiological markers. We determined level of contamination with Legionella of cooling tower in Seoul city, analyzed the distribution of Legionella subtypes, and evaluated molecular typing methods for discrimination power and feasibility.
METHODS
Water samples from 120 cooling towers in 25 areas(Gu) of Seoul city were collected during June, 1997. Culture and duplex-PCR(polymerase chain reaction) with Southern hybridization probed with Legionella-specific genes were performed with filtered samples. Twenty-two Legionella isolates were analyzed comparatively by pulsed-field gel electrophoresis(PFGE) and arbitrarily primed(AP)-PCR using a M13 reverse primer.
RESULTS
Culture and duplex-PCR with Southern hybridization were positive for Legionella in 22(18.3%) and 106(88.3%) of 120 samples, respectively, resulting in 90.8%(109/120) of contamination level. Out of 22 Legionella isolates, 17 were identified as Legionella pneumophila serogroup 1, 4 as L. pneumophila serogroup 6 and 1 as an unknown. Molecular analysis of 17 isolates of L. pneumophila serogroup 1 showed 7 subtypes by PFGE(A0 in 9 isolates; A1, 2; A2, 1; A3, 2; B, 1; C, 1; D, 1) and 5 subtypes by AP-PCR(Ia in 11 isolates; Ib, 2; Ic, 2; II, 1; III, 1). The agreement of results of both methods was 76.5%(13/17) of L. pneumophila serogroup 1 and 81.8%(18/22) of all isolates, respectively.
CONCLUSION
Most of cooling towers in Seoul city were already contaminated with Legionella just before summer, requiring decontamination measures and continuous surveillance. L. pneumophila serogroup 1 was the predominant isolate with variable subtypes. The AP-PCR can be used as a rapid and reproducible screening tool in tracking legionellosis outbreak.